CN106755179A - A kind of culture medium for being suitable to bacteria cellulose fermentation - Google Patents
A kind of culture medium for being suitable to bacteria cellulose fermentation Download PDFInfo
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- CN106755179A CN106755179A CN201611113691.1A CN201611113691A CN106755179A CN 106755179 A CN106755179 A CN 106755179A CN 201611113691 A CN201611113691 A CN 201611113691A CN 106755179 A CN106755179 A CN 106755179A
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- bacteria cellulose
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- fermentation
- acetobacter
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
Abstract
The invention belongs to fermentation technical field, and in particular to a kind of culture medium for being suitable to bacteria cellulose fermentation.The culture medium is using corn flour, analysis for soybean powder, bean cake powder as major ingredient, not only reduce cost, and raw material it is cheap and easy to get, it is nutritious comprehensively, processing technology it is simple, more conventional fermentation medium for bacterial cellulose cost reduces by 57 times, and the bacteria cellulose fermentation yield that can complete 24g/L and the above was cultivated by 136 144 hours, improve the production efficiency of cellulose.
Description
Technical field
The invention belongs to fermentation technical field, and in particular to a kind of culture medium for being suitable to bacteria cellulose fermentation.
Technical background
Bacteria cellulose is that certain monoid microorganism (bacterium) is referred to as through the cellulose of synthesis secretion, many with wood in industrial production
Acetobacter makees the strain of bacteria cellulose fermentation.Used as a kind of novel biomaterial, bacteria cellulose is subject to its special performance
Extensive concern.Bacteria cellulose have high-purity, ultra-fine property, permeable breathable, good biocompatibility, degradability and
Biosynthesis Modulatory character, in medical instruments field, food additives field, field of papermaking, field of textiles and field of Environment Protection
Have broad application prospects.
Microbial fermentation is a complexity, the biological process of system, and period is influenceed by many factors, such as culture medium
Component, proportioning, cultivation temperature, training method.Wherein, the composition of culture medium and proportioning are one of key factors, are entered both at home and abroad
Numerous studies are gone.
Domestic bacteria cellulose production is initial with coconut water as primary raw material, such as the hair of Application No. 201110223884.3
A kind of fermentation medium for bacterial cellulose is disclosed in bright application, is contained in the culture medium and is accounted for culture medium gross weight 30-90%'s
The coconut dregs of rice add water the slurries that obtain after mixing.The method is not easy to wide popularization and application by territory restriction.Scientific research and production people
Member successively develops Juice, plant extract and all kinds of agricultural byproducts leftover bits and pieces etc. as bacteria cellulose fermentation major ingredient.
Current media is primarily present following shortcoming：1) feed preparation process is complicated, expends larger man power and material.2) raw material are received
Territory restriction, is unfavorable for promoting.3) bacteria cellulose yield is relatively low, increased the unit cost of production, high production cost limit
The popularization and application of bacteria cellulose are made, current bacteria cellulose is mainly limited to added value product scope higher.
Shi Chan grains big country of China, possesses abundant cultivated land resource, and corn is the third-largest cereal crops of China, aboundresources,
It is quality-high and inexpensive.Corn contains abundant starch, protein and trace element, is ideal culture medium raw material, such as China
A kind of preparation methods of bacteria cellulose of patent of invention ZL201110110794.3 disclose one kind and add corn in the medium
Core enzymolysis liquid is come the method for preparing bacteria cellulose, the method cellulase, hemicellulase and zytase enzymolysis corn
Core is made corncob enzymolysis liquid, and in the culture medium for adding it to prepare bacteria cellulose, can make what is be rich in corncob
Xylan digests into the xylo-oligosaccharide with good healthcare function, and is wrapped or adsorbs in bacteria cellulose, improves bacterium
The healthcare function of cellulose.But not yet find to produce the technology of bacteria cellulose using corn powder.
Soybean is one of important cereal crops of China, containing rich in protein, the carbon of high-quality can be provided for microorganism
The nutrient raw materials such as source, nitrogen source.Accessory substance -- the dregs of beans of beans processing because containing abundant carbohydrate and protein and
Trace element, is also preferable microbial fermentation raw material.
The report of bacteria cellulose fermentation is applied to currently without corn powder, analysis for soybean powder and dregs of beans.
The content of the invention
Present invention aim at：A kind of applicable culture medium for significantly improving bacteria cellulose output is provided, used by culture medium
Raw material is cheap and easy to get, nutritious comprehensive, processing technology is simple.While cost of material is reduced, bacterium is significantly improved fine
The plain yield of dimension, is greatly lowered the production cost of bacteria cellulose, effectively facilitates the popularization and application of bacteria cellulose.The present invention is used
Corn flour, analysis for soybean powder, bean cake powder not only reduce cost, and comprehensive nutrition as the major ingredient of bacteria cellulose fermented and cultured,
Easily separated with product, be conducive to the volume increase and collection of bacteria cellulose.
The culture medium is specific as follows：
(1) seed culture medium：1000 parts by weight meters, corn flour 5-15 parts, analysis for soybean powder 5-12 parts, white granulated sugar 5-10 parts,
Yeast extract 1-3 parts, tryptone 1-3 parts, 3 parts of potassium dihydrogen phosphate, 1 part of magnesium sulfate, balance of water, to 6.8,121 DEG C go out pH
Bacterium 20 minutes；
(2) fermentation medium：1000 parts by weight meters, corn flour 8-20 parts, bean cake powder 5-20 parts, white granulated sugar 5-15 parts,
2 parts of potassium dihydrogen phosphate, 2 parts of magnesium sulfate, 1 part of calcium chloride, balance of water, to 6.8,121 DEG C sterilize 20 minutes pH；
The corn flour is crushed to 60 mesh and following by whole kernel corn；
The analysis for soybean powder is crushed to 60 mesh and following by soybean grain；
The bean cake powder is a leaching dregs of beans, is crushed to 60 mesh and following；
The white granulated sugar is food grade certified products；
The yeast extract, tryptone, potassium dihydrogen phosphate, magnesium sulfate, calcium chloride are commercially available conventional reagent；
The present invention also provides a kind of method for preparing bacteria cellulose using above-mentioned culture medium fermentation：
(1) by strain after routinely expanding numerous tube, in the above-mentioned seed culture fluid of access；
The strain can be xylose gluconic acid acetobacter (Gluconacetobacter xylinus), acetobacter aceti
(Acetobacter aceti) Pasteur's acetobacter (Acetobacterpasteutinus), Hansen acetobacter
Or tunica albuginea acetobacter (Acetobacter acetosus) (Acetobacterhansenii)；
(2) seed culture condition is：25 DEG C in constant-temperature table, 120-150rpm cultivate 40-48 hour obtain seed liquor；
(3) bacteria cellulose fermenting and producing is using three-stage control：
First stage：Seed liquor is seeded in fermentation medium by the inoculum concentration of 5-10%, 26-28 DEG C, 120-
180rpm is cultivated 24 hours；
Second stage：Cultivation temperature is adjusted to 28-30 DEG C, 150-240rpm is cultivated 48 hours；
Phase III：28-30 DEG C, 80-160rpm continue cultivate 48 hours, obtain bacteria cellulose dynamic fermentation particle；
(4) the bacteria cellulose dynamic fermentation particle for obtaining fermented and cultured soaks through deionized water rinsing, 1%NaOH solution
Boil, deionized water rinsing to neutrality, after freeze-drying target product.
Beneficial effect：
(1) the raw materials used comprehensive, processing technology cheap and easy to get, nutritious of culture medium that the present invention is provided is simple；In drop
While low cost of material, significantly improve bacteria cellulose output to 24g/L and more than, effectively increase bacteria cellulose
Yield, is greatly lowered the production cost of bacteria cellulose, effectively facilitates the popularization and application of bacteria cellulose；
(2) corn flour of the invention, analysis for soybean powder, bean cake powder as bacteria cellulose fermented and cultured major ingredient, it is more conventional thin
5-7 times of fungin fermentation medium cost reduction, and comprehensive nutrition, easily separate with product, are conducive to bacteria cellulose
Cleaning and collection；
(3) not only low cost, yield are high for culture medium provided by the present invention, and improve the production efficiency of cellulose,
The bacteria cellulose fermentation yield of 24g/L and the above can be completed by culture in 136-144 hours.
(4) bacteria cellulose fermentation process of the invention, first using the fermentation process of three-stage temperature-speed-changing, according to bacterium
Inoculum concentration and metabolic alterations regulation temperature and the rotating speed planted, it is easy to operate, effectively increase the yield of bacteria cellulose.
Brief description of the drawings：
Fig. 1：Bacteria cellulose flow sheet
Specific embodiment：
Strain used is xylose gluconic acid acetobacter (Gluconacetobacter xylinus) in following examples
CCTCC AB2040512；
During embodiment fermentation tank dynamic cultivation every 24 hours sampling observation and analytical unit volume biomass once,
Observed using under Gram's stain smear microscopy.
A kind of method for being suitable to the culture medium and fermented-producing bacteria cellulose of bacteria cellulose fermentation of embodiment 1
Seed culture medium：In terms of 1000 parts, 15 parts of corn flour, 5 parts of analysis for soybean powder, 5 parts of white granulated sugar, 1 part of tryptone, yeast
2 parts of medicinal extract, 3 parts of potassium dihydrogen phosphate, 1 part of magnesium sulfate, balance of water, pH is to 6.8 for regulation, and 121 DEG C sterilize 20 minutes；
Fermentation medium：In terms of 1000 parts, 20 parts of corn flour, 6 parts of bean cake powder, 5 parts of white granulated sugar, 2 parts of potassium dihydrogen phosphate, sulphur
2 parts of sour magnesium, 1 part of calcium chloride, balance of water, pH is to 6.8 for regulation, and 121 DEG C sterilize 20 minutes；
The corn flour is crushed to 60 mesh and following by whole kernel corn；The analysis for soybean powder by soybean grain be crushed to 60 mesh and with
Under；The bean cake powder is a leaching dregs of beans, is crushed to 60 mesh and following；The white granulated sugar is food grade certified products；The yeast leaching
Cream, tryptone, potassium dihydrogen phosphate, magnesium sulfate, calcium chloride are commercially available conventional reagent.
It is prepared by seed：Slant strains are connected in seed culture medium in superclean bench, 25 DEG C, 150rpm in constant-temperature table
It is 48 hours of culture, standby.
The bacteria cellulose dynamic fermentation first stage：Seed is seeded in fermentation tank by 10% inoculum concentration, 180rpm,
28 DEG C are cultivated 24 hours；Second stage：By condition of culture be adjusted to 30 DEG C, 210rpm cultivate 48 hours；Phase III：By rotating speed
It is filtered to remove bacterium solution and obtains bacteria cellulose granular materials after is adjusted to 140,28 DEG C of cultures 48 hours.
Product treatment：Gained granular materials is cleaned with deionized water, the hydrogen-oxygen of mass concentration 1% is used after cleaning remaining medium
Change 90 DEG C of sodium solution to process 15 minutes, deionized water rinsing obtains BC (bacteria cellulose) moist wood to neutral, and resulting materials are freezed
Dry BC dry powder, weighs and records.
A kind of method for being suitable to the culture medium and fermented-producing bacteria cellulose of bacteria cellulose fermentation of embodiment 2
Seed culture medium：In terms of 1000 parts, 10 parts of corn flour, 12 parts of analysis for soybean powder, 6 parts of white granulated sugar, 1 part of yeast extract, pancreas
1 part of peptone, 3 parts of potassium dihydrogen phosphate, 1 part of magnesium sulfate, balance of water, pH is to 6.8 for regulation, and 121 DEG C sterilize 20 minutes.
Fermentation medium：In terms of 1000 parts, 14 parts of corn flour, 8 parts of bean cake powder, 10 parts of white granulated sugar, 2 parts of potassium dihydrogen phosphate,
2 parts of magnesium sulfate, 1 part of calcium chloride, balance of water, pH is to 6.8 for regulation, and 121 DEG C sterilize 20 minutes.
It is prepared by seed：Slant strains are connected in seed culture medium in superclean bench, 25 DEG C, 150rpm in constant-temperature table
It is 48 hours of culture, standby.
Bacteria cellulose dynamic fermentation：
Seed is seeded in fermentation tank by 6% inoculum concentration, 140rpm, 27 DEG C cultivate 24 hours；Second stage：Will training
Foster condition be adjusted to 29 DEG C, adjustment of rotational speed be 240rpm cultivate 48 hours；Phase III：It is adjusted to 30 DEG C, rotating speed tune
It is whole cultivated 48 hours for 80rpm after be filtered to remove bacterium solution and obtain bacteria cellulose granular materials.
Other processing modes such as embodiment 1.
A kind of method for being suitable to the culture medium and fermented-producing bacteria cellulose of bacteria cellulose fermentation of embodiment 3
Seed culture medium is prepared：In terms of 1000 parts, 5 parts of corn flour, 10 parts of analysis for soybean powder, 7 parts of white granulated sugar, 2 parts of tryptone,
2 parts of yeast extract, 3 parts of potassium dihydrogen phosphate, 1 part of magnesium sulfate, balance of water, pH is to 6.8 for regulation, and 121 DEG C sterilize 20 minutes.
Fermentation medium is prepared：In terms of 1000 parts, 10 parts of corn flour, 14 parts of bean cake powder, 12 parts of white granulated sugar, potassium dihydrogen phosphate
2 parts, 2 parts of magnesium sulfate, 1 part of calcium chloride, balance of water, pH is to 6.8 for regulation, and 121 DEG C sterilize 20 minutes.
It is prepared by seed：Slant strains are connected in seed culture medium in superclean bench, 25 DEG C, 130rpm in constant-temperature table
It is 40 hours of culture, standby.
The valency of bacteria cellulose dynamic fermentation first section：Seed is seeded in fermentation tank by 5% inoculum concentration, setting speed
It is 120rpm, cultivation temperature is set as that 26 DEG C are cultivated 24 hours；Second stage：It is 180rpm, culture by fermentation tank adjustment of rotational speed
Temperature is adjusted to 28 DEG C and cultivates 48 hours；Phase III：By adjustment of rotational speed 160rpm, 29 DEG C of cultures are filtered to remove bacterium after 48 hours
Liquid obtains bacteria cellulose granular materials.
Other processing modes such as embodiment 1.
A kind of method for being suitable to the culture medium and fermented-producing bacteria cellulose of bacteria cellulose fermentation of embodiment 4
Seed culture medium：In terms of 1000 parts, 11 parts of corn flour, 7 parts of analysis for soybean powder, 10 parts of white granulated sugar, 3 parts of yeast extract, pancreas
3 parts of peptone, 3 parts of potassium dihydrogen phosphate, 1 part of magnesium sulfate, balance of water, pH is to 6.8 for regulation, and 121 DEG C sterilize 20 minutes.
Fermentation medium：In terms of 1000 parts, 8 parts of corn flour, 20 parts of bean cake powder, 15 parts of white granulated sugar, 2 parts of potassium dihydrogen phosphate,
2 parts of magnesium sulfate, 1 part of calcium chloride, balance of water, pH is to 6.8 for regulation, and 121 DEG C sterilize 20 minutes.
It is prepared by seed：Slant strains are connected in seed culture medium in superclean bench, 25 DEG C, 120rpm in constant-temperature table
It is 48 hours of culture, standby.
The bacteria cellulose dynamic fermentation first stage：Seed is seeded in fermentation tank by 8% inoculum concentration, 160rpm,
28 DEG C are cultivated 24 hours；Second stage：By condition of culture be adjusted to 30 DEG C, 150rpm cultivate 48 hours；Phase III：29℃、
110rpm is filtered to remove bacterium solution and obtains bacteria cellulose granular materials after cultivating 48 hours.
Other processing modes such as embodiment 1.
The check experiment 1 of embodiment 5
Seed culture medium：Glucose 20g/L, peptone 5g/L, potassium dihydrogen phosphate 4.3g, magnesium sulfate 2.5g, sodium chloride 3g,
Balance of water, adjusts pH to 6.8；
Fermentation medium：Spontaneous fermentation coconut water 1000ml, sucrose 40g, yeast extract 10g, potassium dihydrogen phosphate 4.3g, sulphur
Sour magnesium 2.5g, sodium chloride 3g, balance of water adjust pH to 6.8.
Other processing modes such as embodiment 1.
The check experiment 2 of embodiment 6
Seed culture medium and fermentation medium are prepared with embodiment 1, and with embodiment 1, fermented and cultured uses 7% to seed culture
Inoculum concentration be seeded in fermentation tank, 30 DEG C, 120rpm is filtered to remove bacterium solution and obtains bacteria cellulose granular material after cultivating 120 hours
Material.
Table 1：Each test group fermentation medium cost
| Group | Embodiment 1 | Embodiment 2 | Embodiment 3 | Embodiment 4 | Embodiment 5 | Embodiment 6 |
| Culture medium cost (unit/L) | 0.262 | 0.275 | 0.337 | 0.317 | 1.421 | 0.262 |
| Yield (dry weight g/L) | 26.73 | 24.88 | 28.69 | 25.66 | 1.95 | 12.31 |
Claims (4)
1. it is a kind of to be suitable to the culture medium that bacteria cellulose ferments, it is characterised in that the culture medium is specific as follows：
(1) seed culture medium：1000 parts by weight meters, corn flour 5-15 parts, analysis for soybean powder 5-12 parts, white granulated sugar 5-10 parts, yeast
Medicinal extract 1-3 parts, tryptone 1-3 parts, 3 parts of potassium dihydrogen phosphate, 1 part of magnesium sulfate, balance of water, pH to 6.8,121 DEG C sterilizing 20
Minute；
(2) fermentation medium：1000 parts by weight meters, corn flour 8-20 parts, bean cake powder 5-20 parts, white granulated sugar 5-15 parts, phosphoric acid
2 parts of potassium dihydrogen, 2 parts of magnesium sulfate, 1 part of calcium chloride, balance of water, to 6.8,121 DEG C sterilize 20 minutes pH.
2. the method that culture medium described in a kind of utilization claim 1 prepares bacteria cellulose, it is characterised in that specific steps are such as
Under：
(1) seed culture：Strain is inoculated in seed culture medium, 25 DEG C in constant-temperature table, 120-150rpm cultivate 40-48 hours；
(2) three-stage fermented and cultured：
First stage：Seed liquor is seeded in fermentation medium by the inoculum concentration of 5-10%, 26-28 DEG C, 120-180rpm trainings
Support 24 hours；
Second stage：Cultivation temperature is adjusted to 28-30 DEG C, 150-240rpm is cultivated 48 hours；
Phase III：28-30 DEG C, 80-160rpm continue cultivate 48 hours, obtain bacteria cellulose dynamic fermentation particle；
(3) by fermented and cultured obtain bacteria cellulose dynamic fermentation particle through deionized water rinsing, 1%NaOH solution digestion,
Deionized water rinsing obtains bacterial cellulose product to neutrality after freeze-drying.
3. the method for preparing bacteria cellulose as claimed in claim 2, it is characterised in that the strain is selected from xylose gluconic acid
Acetobacter (Gluconacetobacter xylinus), acetobacter aceti (Acetobacter aceti) Pasteur's acetobacter
(Acetobacterpasteutinus), Hansen acetobacter (Acetobacterhansenii) or tunica albuginea acetobacter
(Acetobacter acetosus)。
4. application of the culture medium described in claim 1 in fermented-producing bacteria cellulose.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109984193A (en) * | 2018-01-03 | 2019-07-09 | 钟宇光 | A kind of preparation method of fermented type coarse cereals Yoghourt |
| CN109984291A (en) * | 2018-01-03 | 2019-07-09 | 钟宇光 | A kind of preparation method of five cereals various grains convenient congee |
| CN112877384A (en) * | 2021-03-26 | 2021-06-01 | 哈尔滨理工大学 | Preparation method of bacterial cellulose, bacterial cellulose-chitosan composite gel skin-care mask and preparation method thereof |
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| CN102994430A (en) * | 2012-12-08 | 2013-03-27 | 福建卫生职业技术学院 | Bacterial cellulose production strain and application thereof |
| CN104031956A (en) * | 2014-06-05 | 2014-09-10 | 陕西科技大学 | Bacterial cellulose fermentation medium made from apple pomace and method for producing bacterial cellulose by utilizing medium |
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2016
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| CN101319242A (en) * | 2008-06-25 | 2008-12-10 | 许春元 | Method for preparing bacteria cellulose with association of activity and inertia |
| CN102994430A (en) * | 2012-12-08 | 2013-03-27 | 福建卫生职业技术学院 | Bacterial cellulose production strain and application thereof |
| CN104031956A (en) * | 2014-06-05 | 2014-09-10 | 陕西科技大学 | Bacterial cellulose fermentation medium made from apple pomace and method for producing bacterial cellulose by utilizing medium |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109984193A (en) * | 2018-01-03 | 2019-07-09 | 钟宇光 | A kind of preparation method of fermented type coarse cereals Yoghourt |
| CN109984291A (en) * | 2018-01-03 | 2019-07-09 | 钟宇光 | A kind of preparation method of five cereals various grains convenient congee |
| CN112877384A (en) * | 2021-03-26 | 2021-06-01 | 哈尔滨理工大学 | Preparation method of bacterial cellulose, bacterial cellulose-chitosan composite gel skin-care mask and preparation method thereof |
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| CN106755179B (en) | 2018-10-30 |
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